pcDNA3 PBD HK-AA (Nigg AH13)
(Plasmid #41161)

• Depositing Lab: Erich Nigg
• Publication: Hanisch et al Mol Biol Cell. 2006 Jan;17(1):448-59. Epub 2005 Nov 2.

Backbone

• Vector backbone: pcDNA3.1/3x myc-C
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5500
• Total vector size (bp): 6331
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: PBD HK-AA
• Alt name: Plk1-PBD HK-AA
• Alt name: Plk1 Polo Box Domain
• Alt name: Polo-like kinase 1 Polo Box Domain
• Species: H. sapiens (human)
• Insert Size (bp): 831
• Mutation: Contains Plk1-PBD only (aa 326–603); HK-AA (H538A, K540A)
• GenBank ID: NM_005030.3 X73458.1
• Entrez Gene: PLK1 (a.k.a. PLK, STPK13)
• Promoter: CMV
• Tags / Fusion Proteins:
  • 3xMyc (N terminal on backbone)
  • PreScission Site (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: BGH-rev

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Addgene plasmid #41162: Plk1-PBDWT (aa 326–603) was cloned in-frame by PCR into a pcDNA3.1 vector encoding an amino-terminal 3xmyc-tag (Invitrogen, Carlsbad, CA) using primers 5
'-CCG GAA TTC CAG ATC TTC GAT TGC TCC CAG CAG CCT GG-3
' and
5
'-CCG CTC GAG TTA GGA GGC CTT GAG ACG GTT GC-3
'.

The Plk1-PBDAA mutant (H538A, K540A) was made by site-directed mutagenesis using the following primers: 5'
-CAA TTC TCC GGA GCC CCG CCT ATC TGT CCC-3
' and 5
'-GGG ACA GAT AGG CGG GGC TCC GGA GAA TTG-3
'.